Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
  • C12N2710/00011—Details
  • C12N2710/16011—Herpesviridae
  • C12N2710/16611—Simplexvirus, e.g. human herpesvirus 1, 2
  • C12N2710/16622—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

• HSV Herpes simplex viruses
• HSV DNA polymerase The major effort to develop antiherpetic drugs has historically centered on nucleoside analog inhibitors of HSV DNA polymerase. All currently used therapies are nucleoside analogs, acyclovir being the prime example. Oral or IV acyclovir is the therapy of choice for most infections. Topical acyclovir, vidarabine or idoxuridine are all used for herpes keratitis, the leading cause of corneal blindness in this country.
• HSV ribonucleotide reductase (RR) is one such target; the viral-specified enzyme is markedly different from mammalian counterparts.
• HSV-RR catalyzes the reduction of the four ribonucleotides to the corresponding deoxyribonucleotides required for DNA replication.
• Herpes RR inhibitors have been shown to possess antiherpetic activity per se (Shipman et al ., Antiviral Research , 6 : 197 (1986)) and also to potentiate or synergize the action of acyclonucleoside antiviral agents (Spector et al ., Proc. of the Nat. Acad. of Sci. , 86 : 1051 (1989)).
• Dutia et al. Nature 321 : 439-441 (1986) and Cohen et al ., Nature 321 : 441-443 (1986) and U.S. Patent No. 4,795,740, both disclosed that the nonapeptide Tyr Ala Gly Ala Val Val Asn Asp Leu, inhibited in vitro the activity of this enzyme.
• Dutia et al ., op . cit . also disclosed that its 8-desalanine homolog, Tyr Gly Ala Val Val Asn Asp Leu, also inhibited in vitro the activity of this enzyme. Gaudreau et al ., J. Biol.
• a series of substituted peptides have been found to inhibit the activity of the ribonucleotide reductase enzyme of herpes simplex virus in vitro .
• the present invention provides novel substituted peptide compounds of the Formula I: wherein:
• A1 and A2 are independently:
• A3 is wherein:
• R4 is hydrogen or methyl
• R5 is -OR8 or NR6R7
• R6 and R7 are independently:
• A4 is aspartic acid, N-methyl aspartic acid, or any of the enantiomorphic forms thereof;
• A5 is an amino acid residue of the formula: wherein:
• R4 is as described hereinabove
• R9 and R10 are independently:
• R a is wherein:
• R1 is H or C1-C6 alkyl
• R2 is:
• R3 is:
• m 0 or 1.
• n 1 if m is 0 or n is 2 if m is 1.
• R4 is hydrogen
• R5 is -NR6R7
• R6 is hydrogen
• R7 is hydrogen or benzyl
• A4 is aspartic acid
• A5 is leucine and m is 0, then R2 is not C1-C4 alkyl monosubstituted by one of the substituents w)-aa).
• the present invention also provides for the pharmaceutically acceptable salts of the above compounds.
• alkyl, alkenyl and alkynyl are intended to include linear and branched structures.
• alkyl is intended to include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl and the like.
• alkenyl is intended to include vinyl, allyl, isopropenyl, pentenyl, hexenyl and the like.
• alkynyl is intended to include ethynyl, propynyl, butynyl and the like.
• cycloalkyl is intended to include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
• cycloalkenyl is intended to include cyclopentenyl, cyclohexenyl, cycloheptenyl and the like.
• heteromatic ring system is intended to include pyridine, thiophene, furan and the like.
• heteromatic polycyclic ring system is intended to include quinoline, isoquinoline, indole, benzofuran, benzothiophene and the like.
• aromatic polycyclic ring system is intended to include naphthalene, phenanthrene and the like.
• halogen is intended to include fluorine, chlorine, bromine or iodine atom.
• Some of the compounds described herein contain one or more centers of asymmetry and may thus give rise to diastereoisomers and optical isomers.
• the present invention is meant to comprehend such possible diastereomers as well as their racemic and resolved optically active forms.
• A3 is an amino acid residue having the formula: where R4 is hydrogen and R5 is -NR6R7 where R6 and R7 are independently hydrogen, methyl, ethyl, benzyl, or R6 and R7 are combined to form a C3-C5 diradical or -NR6R7 is morpholino.
• A5 is an amino acid residue having the formula: where R4 and R9 are hydrogen, and R10 is t-butyl or isopropyl.
• R1 is hydrogen.
• R2 is phenyl, benzyl, or phenyl or benzyl substituted by -NH2, C1-C4 alkyl, -SR8, CO2H or OR8; where R8 is H or C1-C4 alkyl.
• One aspect of this invention involves a pharmaceutical composition
• a pharmaceutical composition comprising an antiherpes virally effective amount of a compound of Formula I, or a therapeutically acceptable salt thereof, and a pharmaceutically or veterinarily acceptable carrier.
• Another aspect of this invention involves a method of treating herpes viral infection in a mammal by administering to the mammal an antiherpes virally effective amount of the peptide of Formula I or a therapeutically acceptable salt thereof as defined hereinafter.
• Another aspect of this invention involves a method of treating viral infections in mammals comprising administering to the mammal an antivirally effective amount of the peptide of Formula I with another antiviral acyclonucleoside or a related compound.
• the peptides of the instant invention and amides and salts thereof can be manufactured according to known synthetic methods, i.e., by condensing amino acids stepwise or by solid phase synthesis according to the method originally described by Merrifield, J. Am. Chem. Soc. , 85 : 2149-2154 (1963) or by using automated peptide synthesizing equipment.
• the N-substituent can then be attached to the pentapeptide unit according to known synthetic methods, i.e., by reaction with the corresponding activated acylating agent in the presence of an organic nitrogen base.
• activated acylating agent may be acid chloride, acid anhydride, pentafluorophenyl ester and carbamyl chloride.
• the condensation between two amino acids can be carried out according to the usual condensation methods such as azide method, mixed acid anhydride method, DCC (dicyclohexyl carbodiimide) method, N-hydroxysuccinimide method, cyano method, Woodward reagent K method, carbonyl diimidazole method or oxidation reduction method. These condensation reactions may be done in either liquid or solid phase. In the case of elongating the peptide chain in the solid phase method, the peptide is attached to an insoluble carrier at the C-terminal amino acid.
• condensation methods such as azide method, mixed acid anhydride method, DCC (dicyclohexyl carbodiimide) method, N-hydroxysuccinimide method, cyano method, Woodward reagent K method, carbonyl diimidazole method or oxidation reduction method.
• halomethyl resin such as chloromethyl resin and bromomethyl resin, benzhydrylamine resin, and t-alkoxy carbonyl hydrazide resin can be used.
• the applicable protective groups to amino groups are exemplified such as benzyloxycarbonyl (hereinafter abbreviated as Z), o-chlorobenzyloxy carbonyl [Z(2-C1)], p-nitrobenzyloxy carbonyl [Z(NO2)], p-methoxybenzyloxycarbonyl [Z(-OMe)], t-butoxycarbonyl (BOC), t-amyloxycarbonyl (Aoc), isobornyloxycarbonyl (Bpoc), 9-fluorenylmethoxycarbonyl (Fmoc), methylsulfonyl ethoxy carbonyl (Msc), trifluoroacetyl, phthalyl, formyl, 2-nitrophenyl sulphenyl (NPS), diphenyl phosphin
• protective groups for the carboxy group there can be exemplified, for example, benzyl ester (OBzl), 4-nitrobenzyl ester [OBzl(NO2)], t-butyl ester (OBut), 4-pyridyl methyl ester (OPic), and the like. It is desirable that specific amino acids such as arginine, cysteine, and serine possessing a functional groups other than amino and carboxyl groups are protected by a suitable protective group as occasion demands.
• the guanidino group in arginine may be protected with nitro, p-toluene sulfonyl, benzyloxycarbonyl, adamantyloxycarbonyl, p-methoxybenzenesulfonyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), 4-methoxy-2,6-dimethyl benzenesulfonyl (Mds), 1,3,5-trimethylphenylsulfonyl (Mts), and the like.
• the thiol group in cysteine may be protected with benzyl, p-methoxybenzyl, triphenylmethyl, acetylaminomethyl, ethyl carbamoyl, 4-methylbenzyl, 2,4,6-trimethylbenzyl (Tmb), etc, and the hydroxy group in serine can be protected with benzyl, t-butyl, acetyl, tetrahydropyranyl, etc.
• compositions of the present invention comprise a compound of Formula I as an active ingredient or a pharmaceutically acceptable salt, thereof, and may also contain a pharmaceutically acceptable carrier and optionally other therapeutic ingredients.
• pharmaceutically acceptable salts refers to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts.
• Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N1-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.
• basic ion exchange resins such as argin
• salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids.
• acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like.
• Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids.
• a pharmaceutical composition or preparation comprising a compound of the Formula I as hereinbefore defined; or a therapeutically acceptable salt thereof, together with a pharmaceutically acceptable carrier therefore.
• the pharmaceutical composition comprises a compound of the present invention in effective unit dosage form.
• an effective unit dosage or “effective unit dose” is denoted to mean a predetermined antiviral amount sufficient to be effective against the viral organisms in vivo .
• Pharmaceutically-acceptable carriers are materials useful for the purpose of administering the medicament, and may be solid, liquid, or gaseous materials, which are otherwise inert and medically acceptable and are compatible with the active ingredients.
• compositions may be given parenterally, orally, used as a suppository or pessary, applied topically as an ointment, cream, aerosol, powder, or given as eye or nose drops, etc., depending on whether the preparation is used to treat internal or external viral infections.
• compositions are administered orally or parenterally at dose levels of about 0.1 to 250 mg per kg, preferably 1.0 to 50 mg per kg of mammal body weight, and are used in man in a unit dosage form, administered, e.g. a few times daily, in the amount of 1 to 250 mg per unit dose.
• fine powders or granules may contain diluting, dispersing and/or surface active agents, and may be presented in a draught, in water or in a syrup; in capsules or sachets in the dry state or in a non-aqueous solution or suspension, wherein suspending agents may be included; in tablets, wherein binders and lubricants may be included; or in a suspension in water or a syrup.
• suspending agents may be included
• binders and lubricants may be included
• a suspension in water or a syrup where desirable or necessary, flavoring, preserving, suspending, thickening, or emulsifying agents may be included. Tablets and granules are preferred, and these may be coated.
• the compounds may be presented in aqueous solution in a concentration of from about 0.1 to 10%, more preferably 0.1 to 7%, most preferably 0.2% w/v.
• the solution may contain antioxidants, buffers, etc.
• compositions are preferably applied to the infected part of the body of the patient as a topical ointment or cream.
• the compound may be presented in an ointment, for instance, with a water soluble ointment base, or in a cream, for instance with an oil in a water cream base, in a concentration of from about 0.1 to 10%, preferably 0.1 to 7%, most preferably 1% w/v.
• the title compound was synthesized by using a Milligen 9050 automated peptide synthesizing machine employing the commercially available PepSyn KA resin and the corresponding single peptide units commercially available as the N-9-fluorenylmethoxycarbonyl-pentafluorophenyl esters.
• the polypeptide-resin product from the synthesizer was treated with 95: 4: 1 trifluoroacetic acid: 1,2-ethanedithiol: thioanisole, followed by aqueous 80% acetic acid.
• the crude product in solution was then purified by reverse phase HPLC to provide the title compound. p.m.r.
• Step A N-Carbobenzyloxyaspartic acid ⁇ -Benzyl- ⁇ -p-nitrophenyl diester
• Step B N-Carbobenzyloxy-N′,N′-dimethylasparagine benzyl ester
• Step C N′,N′-Dimethylasparagine
• Step D N-(Fluorenylmethoxycarbonyl)-N′,N′-dimethyl-asparagine
• the layers were separated and the aqueous phase was washed 2 times with ethyl ether.
• the combined organic phases were back extracted with water and the combined aqueous phases were acidified to pH 2.
• the aqueous mixture was then extracted with ethyl ether (3 ⁇ 60 mL) and the combined organic phases were dried over anhydrous sodium sulfate.
• the solution was then concentrated under vacuum to provide the title compound which was used as is in a subsequent reaction.
• Step E N-(Fluorenylmethoxycarbonyl)-N′,N′-dimethyl-asparagine pentafluorophenyl ester
• the substituted aparagine of Step D (545 mg) was treated with 263 mg of pentafluorophenol and the mixture dissolved in 10 mL of ethyl acetate. The solution was cooled to 0°C and solution of 295 mg of DCC in 5 mL of ethyl acetate was added. The mixture was stirred for 45 minutes at 0°C and for 75 minutes at RT. The mixture was filtered through a pad of Celite and concentrated under vacuum. Flash chromatography of the residue over silica gel (1: 1 ethyl acetate: hexane) provided the title compound.
• Step F L-Aspartyl-L-valyl-L-valyl-L-(N′,N′-dimethyl-asparagyl)-L-aspartyl-L-leucine
• Step A 2-Benzyl-2-(4-methoxybenzyl)acetic acid
• Step B D,L-2-Benzyl-2-(4-methoxybenzyl)acetic acid pentafluorophenyl ester
• Step D D-N-(2-Benzyl-2-(4-methoxybenzyl)-acetyl)- L-valyl-L-valyl-L-asparagyl-L-aspartyl- L-leucine and L-N-(2-Benzyl-2-(4-methoxybenzyl)-acetyl)- L-valyl-L-valyl-L-asparagyl-L-aspartyl- L-leucine
• Example 4 Using the procedure of Example 4, but substituting L-valyl-L-valyl-L-(N′,N′-dimethyl-asparagyl-L-aspartyl-L-leucyl PepSyn KA resin for the Merrifield resin bound peptide in Example 4 provided the title compound.
• p.m.r. CD3CO2D ⁇ : 0.85-1.05 (m, 21H), 1.63 (m, 1H), 1.85 (broad s, 2H), 2.05 (m, 2H), 2.70-3.10 (m, 15H), 4.20 (m, 2H), 4.62 (dd, 1H), 4.98 (m, 2H), 7.18 ppm (m, 10H); M.S. (FAB): m / e 823 (M+H)+.
• Step A t-Butyl p-t-butoxyhydrocinnamate
• Step B p-t-Butoxyhydrocinnamic acid
• Step C p-t-Butoxyhydrocinnamic acid pentafluorophenyl ester
• Step D N-(p-Hydroxyhydrocinnamyl)-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step A 4-(t-Butyldimethylsiloxy)benzoic acid
• Step B 4-(t-Butyldimethylsiloxy)benzyl alcohol
• Step C 4-(t-Butyldimethylsiloxy)benzyl chloride
• Step D p-(t-Butyldimethylsiloxy)hydrocinnamic acid
• Step E 2,2-Bis(p-(t-butyldimethylsiloxy)benzyl)-acetic acid
• a solution of 0.673 mL of diisopropylamine in 3.0 mL of anhydrous THF was cooled to 0°C and the solution treated with 1.72 mL of a 2.4 M n-butyl-lithium in hexane solution.
• the solution was stirred at 0°C for 45 minutes and then a solution of 448.5 mg of the acid from Step D in 2.5 mL of anhydrous THF was added, followed by 0.317 mL of HMPA.
• the solution was stirred for 1 hour at RT, then the solution was cooled to-20°C and 410 mg of p-(t-butyldimethylsiloxy)benzyl chloride was added.
• Step F 2,2-Bis(p-(t-butyldimethylsiloxy)benzyl)-acetic acid pentafluorophenyl ester
• Step G N-[2,2-Bis(4-hydroxybenzyl)acetyl]-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step B N-(2,2-Dibenzylacetyl)-L-valyl-L-valyl-L-asparagyl-L-(O-t-butyl)aspartyl-L-( ⁇ -methylleucyl) PepSyn KA resin
• Step C N-(2,2-Dibenzylacetyl)-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-( ⁇ -methylleucine)
• the resin bound peptide of Step B was suspended in 4 mL of a 95: 4: 1 trifluoroacetic acid: ethanedithiol: thioanisole and the mixture gently stirred for 2 hours. The mixture was then filtered and the solid stirred with 4 mL of the same solution for 5 minutes. The mixture was again filtered and the solid then extracted with 4 mL of 80% aqueous acid. The combined extracts were concentrated under vacuum and the residue was extracted 4x with ether. HPLC purification of the insoluble material provided the title compound. p.m.r.
• Step B N′-(9-Fluorenylmethoxycarbonyl)-N′-benzyl-asparagine
• the phases were separated and the thick aqueous phase was washed 3x with ether.
• the aqueous phase was then carefully acidified to pH 2 by the slow addition of 2N aqueous HC1.
• the mixture was then filtered and the solid dried under vacuum to provide the title compound as a colorless powder; m.p.: 176-178°C.
• Step C N′-(9-Fluorenylmethoxycarbonyl)-N′-benzyl-asparagine pentafluorophenyl ester
• Step D L-Valyl-L-valyl-L-(N′-benzylasparagyl)-L-aspartyl-L-( ⁇ -methylleucyl) PepSyn KA resin
• the title compound was synthesized using a Milligen 9050 automated peptide synthesizing machine employing the commercially available resin, the corresponding single peptide units commercially available as N-9-fluorenylmethoxycarbonyl-pentafluorophenyl esters and the ester from Step C.
• Step E N-(2,2-Dibenzylacetyl)-L-valyl-L-valyl-L-(N′-benzylasparagyl)-L-aspartyl-L-( ⁇ -methyl-leucine)
• Step A D,L-N-(9-Fluorenylmethoxycarbonyl)- ⁇ -methylphenylalanine pentafluorophenyl ester
• Step B N-(9-Fluorenylmethoxycarbonyl)-D- ⁇ -methylphenylalanyl-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine and N-(9-Fluorenylmethoxycarbonyl)-L- ⁇ -methylphenylalanyl-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step A D,L-N-(9-Fluorenylmethoxycarbonyl)- ⁇ -methyltyrosine pentafluorophenyl ester
• Step B D,L-[N-(9-Fluorenylmethyoxycarbonyl)- ⁇ -methyltyrosyl]-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step C D,L-( ⁇ -Methyltyrosyl)-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• a 1N aqueous sodium hydroxide (NaOH) solution (0.055 mL) was added to a slurry of 20.0 mg of L-tyrosyl-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine (synthesized using a Milligen 9050 automated peptide synthesizing machine) in 0.165 mL of water.
• the cloudy solution was diluted with 0.22 mL of acetonitrile and then a solution of 20.2 mg of biphenyl-4-carboxaldehyde in 0.2 mL of acetonitrile was added.
• Step A N-(9-Fluorenylmethoxycarbonyl)-N-methyl-L-leucine
• Step B N-(9-Fluorenylmethoxycarbonyl)-N-methyl-L-leucyl anhydride
• Step C N-(9-Fluorenylmethoxycarbonyl)-N-methyl-L-leucyl PepSyn KA resin
• Step D L-Tyrosyl-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-(N-methylleucine)
• Step B L-[N-(Dibenzylcarbamyl)valyl]-L-valyl-L-asparagyl-L-aspartyl-L-leucyl-PepSyn KA resin
• Step C L-[N-Dibenzylcarbamyl)valyl]-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step C Using the procedure of Example 8, Step C, but substituting the resin bound peptide of Step B for the resin bound peptide of Example 8. Step C, provided the title compound. p.m.r. (CD3OD) ⁇ : 0.77 (d, 3H), 0.88 (d, 3H), 0.95 (d, 3H), 1.0 (m, 9H), 1.65 (m, 1H), 1.80 (m, 2H), 2.04 (m, 1H), 2.14 (m, 1H), 2.70 (dd, 1H), 2.80-294 (m, 3H), 4.17 (t, 2H), 4.40-4.55 (m, 3H), 4.66-4.76 (m, 3H), 4.80 (t, 1H) 7.30 (m, 6H), 7.36 (m, 4H), 7.98-8.10 ppm (m, 1H); M.S. (FAB): m / e 782 (M+H)+.
• Step A N-(Fluorenylmethoxycarbonyl)-N-benzyl-L-(4-t-butoxyphenylalanine)
• Step B L-(N-Benzyltyrosyl)-L-valyl-L-valyl-L-(N′,N′-dimethylasparagyl)-L-aspartyl-L-(y-methylleucine
• Step A Dimethyl-2,2-bis(1-naphthylmethyl)malonate
• Step B 2,2-Bis(1-naphthylmethyl)acetic acid
• Step C 2,2-Bis(1-naphthylmethyl)acetyl chloride
• Step D L-[N-Bis(1-naphthylmethyl)acetylvalyl]-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Step A L-(3,5-Diiodotyrosyl)-L-valyl-L-valyl-L-asparagyl-L-(O-t-butyl)aspartyl-L-leucyl PepSyn KA resin
• the mixture was diluted with 0.9 mL of DMF and swirled 24 hours at RT.
• the mixture was then treated with 2 mL of 20% v/v piperidine/DMF solution and the mixture swirled for 15 minutes.
• the liquid was decanted and the solid treated with 3 mL of the piperidine/DMF solution and swirled for 15 minutes.
• the liquid was again decanted and the solid washed 4x with DMF and 3x with methylene chloride to provide the title compound.
• Step B L-(3,5-Diiodotyrosyl)-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-leucine
• Example 20 Using the procedure of Example 20, but substituting N-(fluorenylmethoxycarbonyl)-N′-benzyl-oxycarbonyllysine for the derivatized tyrosine employed in Example 20, provided the title compound.
• p.m.r (CD3OD) ⁇ : 1.00 (m, 18H), 1.40-2.20 (m, 11H), 2.75 (m, 1H), 2.89 (m, 2H), 3.05 (m, 1H), 3.16 (m, 2H), 4.02 (m, 1H), 4.27 (m, 2H), 4.45 (m, 1H), 4.74 (m, 1H), 4.82 (broad t, 1H), 5.11 (s, 2H), 7.38 (m, 5H), 7.70-8.35 ppm (m, 4H); M.S. (FAB): m / e 821 (M+H)+.
• L-Norvaline (11.72 g) was placed in a large quartz reactor vessel and the vessel cooled to -80°C and 100 ml of anhydrous hydrofluoric acid was condensed inside.
• the vessel was irradiated with UV light and trifluoromethylhypofluorite was bubbled into the vessel for 5 hours. A total of 80 psig of the hypofluorite was used.
• the UV irradiation was continued for an additional 45 minutes, then the solution was stored overnight at -80°C.
• the reaction solution was then concentrated under a stream of nitrogen and the residue was then dissolved in 2 N aqueous HC1 soltuion. This aqueous solution was concentrated under vacuum. HPLC purification of the residue provided the hydrochloride salt of the title compound as well as 4-fluoronorvaline hydrochloride.
• the hydrochloride salt of 5-fluoronorvaline was dissolved in 10 mL of water and the solution filtered thru a Celite plug. The solution was then diluted with isopropyl alcohol (iPrOH) to 75 mL volume and 2 mL of propylene oxide was added. The solution was stirred until the free amino acid began to crystallize out and then it was allowed to stand for 30 minutes. The mixture was diluted with 50 mL of iPrOH and stored in the refrigerator overnight. The mixture was then filtered and the crystals washed with iPrOH and ether. The crystals were then dried under vacuum to provide the title compound.
• iPrOH isopropyl alcohol
• Step B L-Tyrosyl-L-valyl-L-valyl-L-asparagyl-L-aspartyl-L-(5-fluoronorvaline)

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• 1990
  • 1990-11-29 JP JP2326192A patent/JPH03181498A/ja active Pending
  • 1990-12-04 EP EP90313148A patent/EP0438873A1/de not_active Withdrawn
  • 1990-12-06 CA CA002031644A patent/CA2031644A1/en not_active Abandoned

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